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The purpose of this research was to develop a better understanding of the dynamic effects of anaerobic hydraulic retention time (HRT) on both enhanced biological phosphorus removal (EBPR) performance and enrichment of phosphorus accumulating organisms (PAOs). The research was conducted using laboratory-scale sequencing batch reactors inoculated with mixed microbial consortia and fed real wastewater. Exposing microorganisms to extended anaerobic HRTs is not recommended for EBPR configured systems. In this research, however, longer anaerobic exposure did not negatively affect performance even if volatile fatty acids were depleted. Further, extended anaerobic HRTs may positively affect phosphorus removal through enhanced aerobic uptake. The EBPR consortia also appear to maintain reserve energetic capacity in the form of polyphosphate that can be used to survive and grow under variable operational and environmental conditions. Finally, the tested EBPR systems yield mixed microbial consortia enriched with PAOs (specifically Candidatus Accumulibacter phosphatis) at approximately 7.1 to 21.6% of the total population.

In disinfection systems, incomplete penetration of chlorine into effluent wastewater particles can result in a residual population of viable microorganisms. In this work, a combined experimental and numerical approach was used to quantify inactivation of microorganisms in effluent particles and identify combinations of particle removal and chlorine dose that would result in a reduction of occluded microorganisms for six full-scale facilities in the United States with different nitrification levels. The results reveal that combined chlorine is more effective for inactivating occluded microorganisms than free chlorine; model calibration results suggest that free chlorine is less effective because it is more reactive. However, nitrified effluents appear to have lower effluent particle concentrations, and decreases in particle concentrations significantly reduce the chlorine required. Additionally, in disinfection systems that are designed and operated based on inactivation of indicator organisms, the chlorine dose may be insufficient to inactivate occluded pathogens to levels consistent with current regulations.

The influence of four main process parameters--solids retention time (SRT), hydraulic retention time (HRT), anoxic-oxic cycling, and carbon-to-nitrogen ratio (C/N ratio)--on poly(hydroxyalkanoate) (PHA) production, while treating brewery wastewater, was studied. Two sets of two-level, three-factor experimental designs were implemented to (1) determine the effects and interactions among process parameters, (2) assess their significance to PHA production, and (3) approximate optimal operational conditions. The HRT and SRT were found to be the crucial operational parameters affecting PHA production. The highest PHA content of 55% (on a cell-weight basis) was produced at a 4-day HRT and 4-day SRT, whereas a maximum PHA concentration of 907 mg/L was obtained at a 2-day HRT and 12-day SRT. The effect of anoxic conditions on PHA production was insignificant. The C/N ratio played a more important role in the PHA concentration in the system than in the PHA content in the biomass.

The purpose of the research presented herein was to evaluate the effects of solids residence time (SRT) and organic acid augmentation on biological phosphorus removal (BPR), with a focus on how these operational variables affect key metabolisms and the distribution of the microbial population. Using laboratory-scale sequencing batch reactors seeded with a mixed microbial consortium and fed real wastewater, we observed that longer SRTs can improve BPR performance; organic acid augmentation can stabilize BPR, but it is not necessary for process success; and higher volatile suspended solids concentrations correlate with improved phosphorus removal. The results also suggest that organic acids may not be critical in driving anaerobic phosphorus release, but in driving aerobic growth. Finally, given an observed population similarity across all tested bioreactors, BPR variability appears to be less influenced by the presence of specific microbes and more affected by the induction of critical metabolisms.

Polyhydroxyalkanoate (PHA) production was achieved using tomato cannery waste coupled with a mixed microbial culture during wastewater treatment. The two-stage PHA production process comprised a sequencing batch reactor (SBR), operating under a periodic feast-famine regime, to accomplish simultaneously wastewater treatment and selection of PHA-accumulating microbes, followed by a batch reactor for the production of PHA-rich biomass. The SBRs were efficient at removing soluble carbon (84%), ammonia (100%), and phosphorus (76%). Meanwhile, PHA-accumulating microbes were enriched under the SBR operating conditions, and PHA content on a cell-weight basis was within the range 7 to 11% in nonfiltered wastewater and 2 to 8% in filtered wastewater. Subsequently, batch studies were implemented with varying loading rates, ranging from 0.4 to 3.2 food-to-microorganism ratios. A maximum 20% PHA content on a cell-weight basis was obtained. Based on the experimental results, a PHA biosynthesis-degradation kinetic model was developed to (1) aid in the design of a pilot- or full-scale PHA production process coupled with wastewater treatment and (2) determine optimal conditions for harvest of PHA-rich biomass.

Biologically derived polyesters known as polyhydroxyalkanoates (PHAs) represent a potentially "sustainable" replacement to fossil-fuel-based thermoplastics. However, current commercial practices that produce PHA with pure microbial cultures grown on renewable, but refined, feedstocks (i.e., glucose) under sterile conditions do not represent a sustainable commodity. Here, we report on PHA production with a mixed microbial consortium indigenous to an activated sludge process on carbon present in municipal wastewaters. Reactors operated under anaerobic/aerobic and aerobic-only mode and fed primary solids fermenter liquor maintained a mixed microbial consortium capable of synthesizing PHA at 10 to 25% (w/w), while reducing soluble COD by approximately 62 to 71%. More critically, an aerobic batch reactor seeded from the anaerobic/aerobic reactor and fed fermenter liquor achieved approximately 53% PHA (w/w). Results presented suggest that environmentally benign production of biodegradable polymers is feasible. We further used PHA-rich biomass to produce a natural fiber-reinforced thermoplastic composite that can be used to offset advanced wastewater treatment costs.

Direct current applied to a tin oxide anode submersed in water has the potential to generate hydroxyl radicals, a strong chemical oxidant. Tin oxide anode systems, which may represent a viable alternative disinfectant, were investigated for the disinfection of secondary effluents. Coliform bacteria in the effluent were effectively inactivated in a bench-scale tin oxide disinfection system. The number of anodes in the tin oxide disinfection system influenced the steady-state concentration of hydroxyl radicals and, consequently, the contact time necessary to achieve a specified dose. The life expectancy of the anodes was greater than 30 days and no appreciable loss of tin was observed over a 5-day period. A preliminary estimate of the capital and operating costs of a pilot- or full-scale tin oxide disinfection system designed to meet a discharge requirement of 23 most probable number/100 mL at a 3 785-m3/d (1-mgd) facility was comparable to UV and chlorination/dechlorination facilities. Based on this preliminary assessment, tin oxide disinfection systems are a promising alternative disinfectant for secondary effluents.

Polymerase chain reaction (PCR) was used to detect Cryptosporidium parvum oocysts in secondary effluent samples collected from activated-sludge facilities. Serial dilutions of the purified nucleic acid extracts from the samples were made and PCR was conducted to estimate the C. parvum oocyst concentration via a Poisson distribution-based most probable number (MPN). The degree of oocysts associated with wastewater particles was also evaluated. The sensitivity of the MPN-PCR assay was 20 oocysts/PCR unit. The detection limit of the concentration, extraction, and purification protocols in phosphate buffer saline spiked with a known concentration of oocysts ranged from 1.1 to 4.6 oocysts/L; the detection limit for the wastewater samples ranged from 11 to 4200 oocysts/L depending on the extent of inhibition in each sample. The recovery efficiency of the oocysts ranged from 48 to 59% in most samples. Oocysts were found in two out of seven samples with concentrations of 203 and 308 oocysts/L, as estimated by the MPN-PCR method. The oocysts were found only in the filtrate of the grab samples; particle-associated oocysts were not detected. Association of spiked C. parvum oocysts with particles in secondary effluent drawn from wastewater plants with varying operating conditions indicated a weak correlation between the degree of association and the mean cell residence time of the system.